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A kind of separation and culture method of Dendrobium officinale suspension cells

A technology of dendrobium candidum and suspension cells, which is applied in the field of plant cell engineering, can solve problems such as the gap between supply and demand, low technical content in product development, and lack of quality standard system

Active Publication Date: 2021-04-13
江苏赛清科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although artificial cultivation has made some progress, the gap between supply and demand is still quite large. The main reasons are that the supply of high-quality seedlings of Dendrobium candidum is insufficient, the lack of quality standard system, the follow-up product development technology content is low, and most of them are at the level of rough processing. The industrialization development of the industry has formed restrictions and impediments to the development momentum

Method used

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  • A kind of separation and culture method of Dendrobium officinale suspension cells
  • A kind of separation and culture method of Dendrobium officinale suspension cells
  • A kind of separation and culture method of Dendrobium officinale suspension cells

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Experimental program
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Effect test

Embodiment 1

[0024] (1) Disinfection: ① Rinse the newborn root tip of Dendrobium candidum with tap water for 30 minutes, place the washed tissue in a sterilized flask that is too heavy for ultra-clean work and sterilize the surface with 75% ethanol for 1 minute, and then sterilize the tissue with sterilized Rinse with distilled water for 3 to 5 times. ② Then disinfect the tissue with 0.5% to 10% sodium hypochlorite for 5 to 10 minutes, discard the disinfectant, and rinse the tissue with sterilized distilled water for 3 to 5 times. ③ Disinfect again with 0.5% to 10% sodium hypochlorite for 3 to 5 minutes, discard the disinfectant, and rinse the tissue with sterilized distilled water for 3 to 5 times.

[0025] (2) Anti-browning: put the sterilized roots of Dendrobium officinale in the browning inhibition medium (Table 1) containing antioxidants, and then shake the flask for about 30 minutes to 1 hour. Then, remove moisture from the tissue using sterile filter paper.

[0026] (3) Separation...

Embodiment 2

[0031] (1) Disinfection: same as embodiment 1 step

[0032](2) anti-browning: same as embodiment 1 step

[0033] (3) Separation of explant cells and improvement of cell telomerase activity: put the root tip of Dendrobium officinale into a sterilized dish containing cutting fluid (Table 2) with antioxidant activity, remove the root cap, and cut from the incision About 1mm as explants, inoculated into WPM pre-medium and cultivated for 30min. Then the tissue was subjected to ultrasonic treatment for 10 min, and the ultrasonic frequency was 35 kHz, so as to increase the activity of tissue cell telomerase. The obtained tissue was inoculated into MS as the basic medium (1% agar, 3% sucrose, 5 mg / L NAA, 0.5 mg / L KT, pH 5.8) for callus induction. Dark cultivation was carried out during the induction process, the cultivation temperature was 25°C±1°C, and the air humidity was 75%±5%.

[0034] (4) Establishment of suspension cell line: After 35 days of culture, the cells with high mer...

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Abstract

A method for isolating and culturing suspension cells of Dendrobium candidum, the method comprising the steps of: separating and culturing root tissue cells of Dendrobium officinale, suspension culture of cells with high merase activity in the quiescent center, and large-scale suspension culture. The invention overcomes the problem that the isolated materials of Dendrobium officinale are easy to brown and cannot be continuously cultivated on a large scale, and the cells with high-end granzyme activity in the quiescent center of the roots are screened out, and the proliferation amount is large under certain conditions, and the cell activity is strong, and the cells are stable all the year round. Large-scale Dendrobium officinale cell suspension culture continuous expansion system.

Description

technical field [0001] The invention belongs to the technical field of plant cell engineering, and in particular relates to a method suitable for the separation and cultivation of dendrobium officinale meristem suspension cell lines. Background technique [0002] Dendrobium officinale is a perennial epiphytic herb of the genus Dendrobium orchidaceae. It is not only a rare and precious Chinese herbal medicine, but also a famous potted flowering plant. The classics of pharmacy "Shen Nong's Materia Medica", "Tao Zang", "Compendium of Materia Medica Supplements", "Pharmacopoeia of the People's Republic of China" (2000-2010 edition) all have records of the medicinal effects of Dendrobium officinale, and praised it as "nourishing yin and tonifying". Treasures". Dendrobium officinale has been listed as a top-grade medicine since the Qin and Han Dynasties. It is a treasure for nourishing yin and nourishing. Its chemical components are diverse. More than 100 chemical components have...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C12N5/02C12N13/00
CPCC12N5/04C12N13/00C12N2509/00C12N2527/00
Inventor 吴冬智强
Owner 江苏赛清科技有限公司